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Method for extracting hirudin from leech saliva

A technology of hirudin and leeches, applied in the direction of leech inhibitors, chemical instruments and methods, specific peptides, etc., can solve the problems of unfavorable development of leech breeding industry, waste of wild leech resources, etc., to improve the intestinal environment and enhance the body's immunity function, the effect of reducing production cost

Inactive Publication Date: 2015-09-23
广西复鑫益生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional acquisition of hirudin is a one-time plunder of leeches, which causes a huge waste of wild resources of leeches, which is not conducive to the development of leech farming.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The method for extracting hirudin from leech saliva of the present invention comprises the following steps:

[0026] A. Put the leech in the container to be fully hungry, put in the inducer until the leech is full, add emetics to make it spit out the saliva in the body, which is the natural hirudin crude product liquid, then take out the living blood-sucking leech and wash it clean before returning to the breeding pool Continue to raise; Collect the hirudin crude product liquid in the container simultaneously, put in the freezer and keep frozen for later use;

[0027] The inducing substance is composed of 10ml of earthworm extract, 5g of chitosan, 1g of astragalus polysaccharide, and 1g of glucose powder;

[0028] Described emetic is the mixed solution of zinc sulfate and ethanol, and its ratio is 1:1, and the weight ratio between the consumption of emetic and leech living body is 1:1;

[0029] B. Thaw the above-mentioned crude hirudin, add 250ml into 4 times the cold ...

Embodiment 2

[0038] The method for extracting hirudin from leech saliva of the present invention comprises the following steps:

[0039] A. Put the leech in the container to be fully hungry, put in the inducer until the leech is full, add emetics to make it spit out the saliva in the body, which is the natural hirudin crude product liquid, then take out the living blood-sucking leech and wash it clean before returning to the breeding pool Continue to raise; Collect the hirudin crude product liquid in the container simultaneously, put in the freezer and keep frozen for later use;

[0040] The inducing substance is composed of 100ml of earthworm extract, 25g of chitosan, 10g of astragalus polysaccharide, and 10g of glucose powder;

[0041] Described emetic is the mixed solution of zinc sulfate and ethanol, and its ratio is 1:1.5, and the weight ratio between the consumption of emetic and leech living body is 1:5;

[0042] B. Thaw the above-mentioned crude hirudin, take 280ml and add it into...

Embodiment 3

[0051] The method for extracting hirudin from leech saliva of the present invention comprises the following steps:

[0052] A. Put the leech in the container to be fully hungry, put in the inducer until the leech is full, add emetics to make it spit out the saliva in the body, which is the natural hirudin crude product liquid, then take out the living blood-sucking leech and wash it clean before returning to the breeding pool Continue to raise; Collect the hirudin crude product liquid in the container simultaneously, put in the freezer and keep frozen for later use;

[0053] The inducing substance is composed of 200ml of earthworm extract, 40g of chitosan, 15g of astragalus polysaccharide, and 20g of glucose powder;

[0054] Described emetic is the mixed solution of zinc sulfate and ethanol, and its ratio is 1:1, and the weight ratio between the consumption of emetic and leech living body is 1:8;

[0055] B. Thaw the above-mentioned crude hirudin, take 290ml and add it into 4...

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PUM

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Abstract

The invention discloses a method for extracting hirudin form leech saliva. The method includes the following steps of a), placing leeches in a container to allow the leeches to be hungered sufficiently, inputting inductive substances to the leeches to allow the same to absorb fully, adding vomitives to allow the leeches to spit the saliva in bodies, taking out blood-sucking leech living bodies to return to a rearing pond to feed continuously, and meanwhile, collecting hirudin crude product liquid in the container; b), freezing above hirudin crude products, adding into precooled cold acetone, stirring prior to placing in a refrigerator for setting overnight, sucking supernatant acetone the next day prior to centrifuging, adding a trichloroacetic acid solution, and centrifuging to remove residues to obtain concentrated liquid; c), eluting the above concentrated liquid with an anion exchange column chromatography method, and performing gel filtration chromatography to obtain finished products. The method has the advantages of reasonable process, convenience and practicability in operation, good quality stability of the finished hirudin products and high yield, one-time pillage on the leeches with a traditional method can be avoided, wild resources of the leeches are protected, and development of leech farming is driven.

Description

technical field [0001] The invention relates to a method for extracting hirudin from leech saliva. Background technique [0002] Hirudin is a protein in leeches, containing 65 amino acid residues and 3 pairs of disulfide bonds. It has highly specific antithrombin activity and inhibits thrombin from binding to the substrate, so it has anticoagulant effect. [0003] The separation and purification of natural hirudin mainly includes the separation and extraction of hirudin from living hirudin and dried leeches. The acquisition of traditional hirudin is a one-time plunder of leeches, which causes a huge waste of wild resources of leeches and is not conducive to the development of leech breeding. Contents of the invention [0004] Aiming at the problems existing in the prior art, the present invention provides a method that has the characteristics of reasonable process, convenient and practical operation, good quality stability and high yield of hirudin finished products, and ...

Claims

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Application Information

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IPC IPC(8): C07K14/815A01K67/033
CPCC07K14/815A01K67/033
Inventor 谢海林
Owner 广西复鑫益生物科技有限公司
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